Membrane Protein Knockdown Cell Line Development

A major approach in the field of mammalian cell biology is the manipulation of the expression of the genes of interest in selected cell lines, to reveal one or several of the gene's function(s) using transient/stable overexpression or knockdown of the gene of interest. The simplest method for RNA interference is the cytosolic delivery of siRNA oligonucleotides, this technique is limited to cells capable of transfection and is primarily utilized during transient in vitro studies. The introduction of shRNA into mammalian cells through infection with viral vectors allows for stable integration of shRNA and long-term knockdown of the targeted gene. Creative Biolabs offers different approaches to knock down the membrane protein of interest.

Membrane Protein Knockdown Cell Line Development with siRNA

Transfection of short interfering RNA oligonucleotides (siRNAs) directly into the cytosol is the simplest approach for RNA interference (RNAi).

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Membrane Protein Knockdown Cell Line Development with shRNA

shRNAs can be delivered into mammalian cells using plasmid and also be given by infecting the cell with virally produced vectors.

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Membrane Protein Knockdown Cell Line Development with siRNA

Small interfering RNA (siRNA) is a double-stranded RNA at first non-coding RNA molecules, similar to miRNA. Transfection of short interfering RNA oligonucleotides (siRNAs) directly into the cytosol is the simplest approach for RNA interference (RNAi). It prevents translation by degrading mRNA after transcription and interfering with the expression of particular genes. As a result, siRNA is frequently utilized in vitro and in vivo to unravel gene function and as a therapeutic agent to suppress disease-related gene expression. Although RNAi remains an important technique in functional genomics and drug discovery, it is still important to choose the right experimental models. In terms of simplicity, flexibility, throughput, and translatability, there is a significant translational gap between in vitro and in vivo models. As a result, novel experimental models that combine the benefits of in vitro and in vivo models are in high demand.

Advantage of Membrane Protein Knockdown Cell Line Development with siRNA

• The simplest approach for RNA interference
• Functionally validated commercial siRNA
• Potentially high level of gene silencing
• Minimal cellular toxicity
• The generation of a short-term cell line with the desired target gene knockdown

Membrane Protein Knockdown Cell Line Development with shRNA

Small hairpin RNAs (shRNA) are made up of two complementary RNA sequences plus a short loop that resembles the hairpin in miRNA. Like siRNAs, shRNAs can be transfected as plasmids expressing shRNAs transcribed by RNA pol III or modified pol II promoters, and also be delivered into mammalian cells by virally produced vectors. shRNAs can integrate with DNA and consist of two complementary 19-22 bp RNA sequences linked by a short loop of 4-11nt, similar to the hairpin found in naturally occurring miRNA.

Advantage of Membrane Protein Knockdown Cell Line Development with shRNA

• Used to generate stable knockdown cell lines
• Greatly increasing reproducibility of results
• Suitable for primary and non-adherent cells
• The only viable technique for untransfectable cells

As a pioneer and the undisputed global leader in stable cell line development, Creative Biolabs offers domestic and international customers the best possible services while at the most competitive price. To learn more about the membrane protein knockdown cell line development service, please feel free to contact us to confirm how we can be involved in your project.